Hi,

Doing some sort of duplication in any study is a good idea. In the case of ELISAs, the assay will usually test each sample in duplicate (or better in triplicate), which will allow you to account for variation within the assay (intra-assay variation) on a particular day. However, running the ELISA again (i.e. duplicating your entire assay on a different day) will allow you to calculate and assess inter-assay variability, or the reproducibility of your results. Preparation of the samples is key in this aspect, and samples should be prepared from stocks where possible to reduce the chance that improper sample prep is impacting on your final results.

Here is an example of how intra-assay work is done

http://www.ncbi.nlm.nih.gov/pubmed/24578473

Hope this helps,

Omar

Hi, we measured our samples always in duplicates. This will provide enough data for statistical validation of the results.

Cheers, Nadine

It's not really required. You should decide this by keeping your major topic into account. For a simple screening with a qualitative result, it's sufficient to run the assay on a single detection. If you need a higher / better precisicion duplicates are helpful. You should know the measurement range of your assay (it's the concentration range with the lowest imprecisicion). If you find reaults outside this range, go for higher (or lower dilutions).

A good ELISA allwas you an imprecisicion below 8%. If you need an higher precision, triplicates or duplicates at different dilutions are helpful.

Hi,

Doing some sort of duplication in any study is a good idea. In the case of ELISAs, the assay will usually test each sample in duplicate (or better in triplicate), which will allow you to account for variation within the assay (intra-assay variation) on a particular day. However, running the ELISA again (i.e. duplicating your entire assay on a different day) will allow you to calculate and assess inter-assay variability, or the reproducibility of your results. Preparation of the samples is key in this aspect, and samples should be prepared from stocks where possible to reduce the chance that improper sample prep is impacting on your final results.

Here is an example of how intra-assay work is done

http://www.ncbi.nlm.nih.gov/pubmed/24578473

Hope this helps,

Omar

I agree with the previous answers that having duplicates or triplicates in any ELISA run is a mainstay. It is important to monitor any consistency/variation within the dilutions of your test samples, between the assays you run and the overall performance of your ELISAs and ensuring that you obtain quality and reliable data at the end of the day. Thanks, Jacinta

Thanks Omar, the paper is what I need.

Repeats are only a mean to know the variation in your assay. If you are developing an assay then it is mandatory to find this variation which is reported as percentage coefficient of variance (%CV) and it must be

I find a link about this issue.

http://www.abcam.com/index.html?pageconfig=resource&rid=12232